Güvercinlerde Çiçek Virusu İzolasyonu

Yıl 2021, Cilt 14, Sayı 4, 444 - 450, 31.12.2021

Mehmet KÜR Oktay KESKİN

https://doi.org/10.30607/kvj.846843

Öz

Bu çalışmada, güvercinlerde yaygın olarak görülen ve en çok kayıp oluşturan hastalıklardan biri olan çiçek virusunun izole edilmesi amaçlandı. Çalışmada, klinik muayenede çiçek nodülü olarak değerlendirilen lezyonlar bulunan 4 farklı güvercin kümesinin her birinden 5 hayvan olmak üzere toplam 20 adet güvercinden nodül örnekleri toplandı. Toplanan doku örneklerinden inokulum hazırlanarak 10 günlük embriyolu tavuk yumurtalarında (ETY) koryoallantoik membrana (CAM) ekimler yapıldı. Yumurtalarda 6-8 gün inkubasyondan sonra CAM üzerinde poklar görüldü ve yine ETY’de 2 kez pasaj yapılarak virusun yoğunluğu artırıldı. Çiçek virusunu doğrulamak amacıyla gerek nodül örneklerinden, gerekse CAM’da görülen pok örneklerinden DNA izolasyonu yapılarak p4b geninin varlığı açısından PCR ile test edildi ve 578 bp’lik bantların görülmesi pozitif olarak değerlendirildi. Güvercinlerde görülen çiçek enfeksiyonlarında hastalardan toplanan nodül örneklerinden güvercin çiçek virusu Türkiye’de ilk kez izole edildi ve etken moleküler olarak doğrulandı.

Anahtar Kelimeler

Çiçek, Güvercin, İzolasyon, PZR

Isolation of Avipoxvirus from Pigeons

Öz

In this study, it was aimed to isolate pox virus which is the causative agent of pox disease which is commonly seen and cause serious lossess in pigeons. In the study, a total of 20 nodule samples from leisons that are clinically which are evaluated as pox nodules clinically were collected. Samples were from four different pigeon flocks each consisting of five samples. Inoculums were prepared from collected tissue samples and inoculated on the chorioallantoic membrane (CAM) of a 10 day old fertilised embryonated chicken eggs. After six to eight days after incubation, pocks were produced by the virus on CAM. Two passages were made further in order to increase the titer of the virus. DNAs were extracted from both nodules and CAM samples and isolated DNAs were amplified by PCR primers for specific for p4b gene. The presence of 578 bp amplified DNA band was regarded as positive in PCR. Nodule samples from infected pigeons were collected and for the first time pox virus from pigeons was isolated and identified molecularly in Turkey. 

Anahtar Kelimeler

Isolation, Pigeon, Pox, PCR

Kaynakça

• Abd El-Samie HA, Mohamed HS, Al-Bakry IM, Manal A. Genomic Characterization of Pigeon Pox Virus in Egypt. Zag Vet J. 2015; 43(1): 94-101.
• Abdallah FM, Hassanin O. Detection and molecular characterization of avipoxviruses isolated from different avian species in Egypt. Virus Genes. 2013; 46:63-70.
• Adebajo MC, Akinyede O, Shittu IA Seroprevalence of fowl pox antibody in indigenous chickens in Jos North and South council areas of Plateau State, Nigeria, Implication for vector vaccine. ISRN Vet Sci, Article. 2012; ID 154971: 4 pages.
• Arda M, Minbay A, Aydın N, Akay Ö, İzgür, M. Çiçek ve Difteri Hastalığı, In: Kanatlı Hayvan Hastalıkları, Üçüncü baskı, Medisan Yayınevi, Ankara, Türkiye. 1997; pp. 163-166.
• Arda M. Poxviridae Familyası, In: Kafes Kuşları Hastalıkları (Kanaryalar, Muhabbet Kuşları, Papağanlar), Ayban Matbaacılık ve Yayıncılık, Ankara, Türkiye. 2007; pp. 188-193.
• Atkinson CT, Wiegand KC, Triglia D, Jarvi ST. Efficacy of Commercial Canarypox Vaccine for Protecting Hawaii Amakihi from Field Isolates of Avipoxvirus. Hawai`i Cooperative Studies Unit Technical Report. 2010; HCSU-01.
• Audarya SD, Riyesh T, Kumar N, Chhabra D, Sikrodia R, Sharda R, Barua S, Garg UK. Molecular Diagnosis of a Cutaneous Form of Pox in Pigeons at Mhow in Madhya Pradesh, India. 2018; IJCMAS, 7(9): 1318-1323.
• Docherty DE, Long RIR, Flickinger EL, Locke LN.. Isolation of poxvirus from debilitating cutaneous lesions on four immature Grackles. Avian Dis. 1991; 35: 244-247.
• Eleazer TH, Harrel JS, Blalock HG. Transmission studies involving a wet fowl pox isolate. Avian Dis. 1983; 27: 542-544.
• Gilhare VR, Hirpurkar SD, Kumar A, Naik SK, Sahu T. Pock forming ability of fowl pox virüs isolated from layer chicken and its adaptation in chicken embryo fibroblast cell culture. Vet World. 2015; 8(3): 245-250.
• Ha HJ, Alley M, Howe L, Gartrell B. Evaluation of the pathogenicity of avipoxvirus strains isolated from wild birds in New Zealand and the efficacy of a fowl pox vaccine in passerines. Vet Microbiol. 2013; 165(3-4): 268-274.
• Jordan FTW, Pattison M. Fowl Pox and Other Avian Poxes, In: Poultry Diseases, Fourth edition, W.B. Saunders Company Ltd., London, England. 1996; pp. 166–172.
• Kabir L, Haque ME, Borty SC, Mustafa K, Kamal MM, Khasruzzaman AKM, Khan SR, Islam MA. Isolation and Molecular Dedection of Fowl Pox and Pigeon Pox Viruses From Recent Outbreak in Bangladesh. Indian J L Sci. 2015; 5(1): 1-7.
• Kulich P, Roubalova E, Dubska L, Sychra O, Šmid B, Literak I. Avipoxvirus in blackcaps (Sylvia atricapilla). Avian Pathol. 2008; 37(1): 101-107.
• Lee LH, Lee KH. Aplication of the polymerase chain reaction for the diagnosis of fowl poxvirus infection. J Virol Methods. 1997; 63: 113-119.
• Luschow D, Hoffmann T, Hafez HM. Differentiation of avian poxvirus strains on the basis of nucleotide sequences of 4b gene fragment. Avian Dis. 2004; 48: 453-462.
• Manorolla G, Pisoni G, Sironi G, Rampin T. Molecular biological characterization of avian poxvirus strains isolated from different avian species. Vet Microbiol. 2010; 140: 1-8.
• Masola SN, Mzula A, Mwega ED, Kasanga CJ, Wambura PN. Detection and Genetic Characterization of an Avipox Virus Isolate from Domestic Pigeon (Columba livia domestica) in Morogoro Region, Eastern Tanzania. AIR. 2015; 3(5): 460-469.
• Offerman K, Carulei O, Gous TA, Douglass N, Willamson AN. Phylogenetic and histological variation in avipoxviruses isolated in South Africa. JGV. 2013; 94: 2338-2351.
• Prukner-Radovcic E, Lüschow D, Grozdanic IC, Tisljar M, Mazija H, Vranesic D.. Isolation and Molecular Biological Investigations of Avian Poxviruses from Chickens, a Turkey and a Pigeon in Croatia. Avian Dis. 2006; 50: 440-444.
• Rahman S, Islam MA, Islam MS, Nazir KHMNH, Khan MSR. Isolation and molecular detection of Avipoxvirus from field outbreaks in Mymensingh, Bangladesh. JAVAR. 2019; 6(1): 54–59.
• Sanchez A, Valdes LM, Rangel LE, Cobos-Marin L. Evaluation of the effect of a pigeon poxvirus autogenous vaccine. Arch Med Vet. 2012; 44: 81-86.
• Tripathy DN, Reed WM. Pox, In: A laboratory manual for the Isolation, Identification and characterization of avian pathogens, Ed; Dufour-Zavala, L., Fifth Edition, American Association of Avian pathologists, New York, USA. 2008; pp. 116-119.
• Tripathy DN. Avipox viruses, In: Virus infection of birds; Poxviridae, Ed; McFerran JB, McNulty MS., Elsevier Sicence Publishers, Amsterdam, Holland. 1993; pp. 5–15.
• Van-Riper C, Forrester DJ. Avian pox, In: Infectious Diseases of Wild Birds, Ed: Thomas N, Hunter DB, Atkinson CT., Black Well Publishing Professional, Ames, Iowa, USA. 2007; pp. 131-176.